title: Antibody immobilization using heterobifunctional crosslinkers.
abstract: Covalent attachment of functional proteins to a solid support is important for biosensors. One method employs thiol-terminal silanes and heterobifunctional crosslinkers such as N-succinimidyl 4-maleimidobutyrate (GMBS) to immobilize proteins through amino groups onto glass, silica, silicon or platinum surfaces. In this report, several heterobifunctional crosslinkers are compared to GMBS for their ability to immobilize active antibodies onto glass cover slips at a high density. Antibodies were immobilized at densities of 74-220 ng/cm2 with high levels of specific antigen binding. Carbohydrate-reactive crosslinkers were also compared to GMBS using a fiber optic biosensor to detect fluorescently-labeled antigen. At the concentrations tested, the antibodies immobilized with carbohydrate-reactive crosslinkers bound more antigen than GMBS immobilized antibodies as indicated by the fluorescence signal.
authors.affiliation: Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375-5348, USA.
authors.forename: L C
authors.initials: LC
authors.lastname: Shriver-Lake
authors.forename: B
authors.initials: B
authors.lastname: Donner
authors.forename: R
authors.initials: R
authors.lastname: Edelstein
authors.forename: K
authors.initials: K
authors.lastname: Breslin
authors.forename: S K
authors.initials: SK
authors.lastname: Bhatia
authors.forename: F S
authors.initials: FS
authors.lastname: Ligler
mesh_terms: D015374:Biosensing Techniques; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005336:Fiber Optic Technology; D005898:Glass; D007074:Immunoglobulin G; D055100:Optical Fibers; D013388:Succinimides
chemical_list: D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D013388:Succinimides; C032138:N-(gamma-maleimidobutyryloxy)succinimide